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BMC Vet Res. 2019 Apr 15;15(1):116. doi: 10.1186/s12917-019-1851-7.

Visual detection of porcine epidemic diarrhea virus using a novel reverse transcription polymerase spiral reaction method.

Author information

1
Henan Provincial Engineering Laboratory of Insects Bio-reactor, China-UK-NYNU-RRes Joint Laboratory of Insect Biology, Nanyang Normal University, Wolong Road 1638, Nanyang, 473061, People's Republic of China.
2
Veterinary Laboratory, Guangzhou Zoo, Guangzhou, 510642, People's Republic of China.
3
College of Animal Science, South China Agricultural University, Guangzhou, 510642, People's Republic of China.
4
Henan Provincial Engineering Laboratory of Insects Bio-reactor, China-UK-NYNU-RRes Joint Laboratory of Insect Biology, Nanyang Normal University, Wolong Road 1638, Nanyang, 473061, People's Republic of China. lunguangyao@163.com.
5
Henan Provincial Engineering Laboratory of Insects Bio-reactor, China-UK-NYNU-RRes Joint Laboratory of Insect Biology, Nanyang Normal University, Wolong Road 1638, Nanyang, 473061, People's Republic of China. jijun020@126.com.

Abstract

BACKGROUND:

Porcine epidemic diarrhea virus (PEDV) is a major etiological agent of porcine epidemic diarrhea around the world. Point-of-care testing in the field is lacking owing to the requirement for a simple, robust field applicable test that does not require professional laboratory equipment. The aim of this study was to establish a novel reverse transcription polymerase spiral reaction (RT-PSR) assay for the rapid detection of porcine epidemic diarrhea virus (PEDV). For the assay, a specific RT-PSR primer pair was designed against a conserved region in PEDV ORF3.

RESULTS:

The RT-PSR was optimized, and PEDV could be detected after a 50 min incubation at 62 °C, in addition to the 15 min required for reverse transcription. No cross-reaction with other porcine infectious viruses was observed. This new method for PEDV detection was 10 times more sensitive than the conventional reverse transcription-polymerase chain reaction (RT-PCR) assay. The positive rates for 65 clinical samples using the new RT-PSR assay and the conventional RT-PCR assay were 58.46% (38/65) and 53.84% (35/65), respectively. In the RT-PSR assay, the addition of a mixture of dyes allowed a positive reaction to be directly observed by the naked eye.

CONCLUSIONS:

These results indicate that this RT-PSR assay is capable of accurately detecting PEDV, and has the advantages of high specificity and sensitivity for the detection of PEDV.

KEYWORDS:

Porcine epidemic diarrhea virus (PEDV); Reverse transcription polymerase spiral reaction (RT-PSR); Sensitivity; Specificity; detection

PMID:
30987635
PMCID:
PMC6466714
DOI:
10.1186/s12917-019-1851-7
[Indexed for MEDLINE]
Free PMC Article

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